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151.
Ajith Anand Emily Wu Zhi Li Sue TeRonde Maren Arling Brian Lenderts Jasdeep S. Mutti William Gordon‐Kamm Todd J. Jones Nicholas Doane Chilcoat 《Plant biotechnology journal》2019,17(8):1636-1645
An efficient Agrobacterium‐mediated site‐specific integration (SSI) technology using the flipase/flipase recognition target (FLP/FRT) system in elite maize inbred lines is described. The system allows precise integration of a single copy of a donor DNA flanked by heterologous FRT sites into a predefined recombinant target line (RTL) containing the corresponding heterologous FRT sites. A promoter‐trap system consisting of a pre‐integrated promoter followed by an FRT site enables efficient selection of events. The efficiency of this system is dependent on several factors including Agrobacterium tumefaciens strain, expression of morphogenic genes Babyboom (Bbm) and Wuschel2 (Wus2) and choice of heterologous FRT pairs. Of the Agrobacterium strains tested, strain AGL1 resulted in higher transformation frequency than strain LBA4404 THY‐ (0.27% vs. 0.05%; per cent of infected embryos producing events). The addition of morphogenic genes increased transformation frequency (2.65% in AGL1; 0.65% in LBA4404 THY‐). Following further optimization, including the choice of FRT pairs, a method was developed that achieved 19%–22.5% transformation frequency. Importantly, >50% of T0 transformants contain the desired full‐length site‐specific insertion. The frequencies reported here establish a new benchmark for generating targeted quality events compatible with commercial product development. 相似文献
152.
Adoptive cell therapy of tumor-infiltrating lymphocytes has shown promise for treatment of refractory melanoma and other solid malignancies; however, challenges to manufacturing have limited its widespread use. Traditional manufacturing efforts were lengthy, cumbersome and used open culture systems. We describe changes in testing and manufacturing that decreased the process cycle time, enhanced the robustness of critical quality attribute testing and facilitated a functionally closed system. These changes have enabled export of the manufacturing process to support multi-center clinical trials. 相似文献
153.
Emily D. Matys Tyler Mackey Christen Grettenberger Elliott Mueller Anne Jungblut Dawn Y. Sumner Ian Hawes Roger E. Summons 《Geobiology》2019,17(5):551-563
Bacteriohopanepolyols (BHPs) are pentacyclic triterpenoid lipids that contribute to the structural integrity and physiology of some bacteria. Because some BHPs originate from specific classes of bacteria, BHPs have potential as taxonomically and environmentally diagnostic biomarkers. For example, a stereoisomer of bacteriohopanetetrol (informally BHT II) has been associated with anaerobic ammonium oxidation (anammox) bacteria and suboxic to anoxic marine environments where anammox is active. As a result, the detection of BHT II in the sedimentary record and fluctuations in the relative abundance of BHT II may inform reconstructions of nitrogen cycling and ocean redox changes through the geological record. However, there are uncertainties concerning the sources of BHT II and whether or not BHT II is produced in abundance in non‐marine environments, both of which are pertinent to interpretations of BHT II signatures in sediments. To address these questions, we investigate the BHP composition of benthic microbial mats from Lake Fryxell, Antarctica. Lake Fryxell is a perennially ice‐covered lake with a sharp oxycline in a density‐stabilized water column. We describe the diversity and abundance of BHPs in benthic microbial mats across a transect from oxic to anoxic conditions. Generally, BHP abundances and diversity vary with the morphologies of microbial mats, which were previously shown to reflect local environmental conditions, such as irradiance and oxygen and sulfide concentrations. BHT II was identified in mats that exist within oxic to anoxic portions of the lake. However, anammox bacteria have yet to be identified in Lake Fryxell. We examine our results in the context of BHPs as biomarkers in modern and ancient environments. 相似文献
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Martin Surbeck Christophe Boesch Catherine Crockford Melissa Emery Thompson Takeshi Furuichi Barbara Fruth Gottfried Hohmann Shintaro Ishizuka Zarin Machanda Martin N. Muller Anne Pusey Tetsuya Sakamaki Nahoko Tokuyama Kara Walker Richard Wrangham Emily Wroblewski Klaus Zuberbühler Linda Vigilant Kevin Langergraber 《Current biology : CB》2019,29(10):R354-R355
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Amanda Linkous Demosthenes Balamatsias Matija Snuderl Lincoln Edwards Ken Miyaguchi Teresa Milner Batsheva Reich Leona Cohen-Gould Andrew Storaska Yasumi Nakayama Emily Schenkein Richa Singhania Stefano Cirigliano Tarig Magdeldin Ying Lin Gouri Nanjangud Kalyani Chadalavada David Pisapia Howard A. Fine 《Cell reports》2019,26(12):3203-3211.e5